KR20190136486A - Extraction apparatus of biochemical materials from biological samples) - Google Patents

Abstract

The present invention relates to an extraction apparatus which is formed in a simple structure, thereby being able to easily carry, quickly perform a genetic test on a collected sample, and be easily used by anyone. The present invention has an effect that can be used quickly, easily, and economically by automating the entire processes.

Description

Extraction apparatus of biochemical materials from biological samples

The present invention relates to an apparatus for extracting a biological material, more specifically, it is formed in a simple structure, which is easy to carry and can perform a genetic test on a collected sample quickly, which can be easily used by anyone, and automates the whole process. The present invention relates to a biological extraction device that can be used quickly and easily and economically.

Recently, with the development of molecular biology and genetic engineering, genetic testing for animals and humans has been introduced and applied. Examples of the substance to be tested include DNA, a messenger RNA encoding a protein, proteins related to them, specific metabolites, and the like. In addition, the purpose of genetic testing includes prediction of risk of developing a genetic disease, identification of carriers, and the like.

Genetic testing and gene amplification are essential for such genetic testing, and the gene extraction starts from the sampling stage.

For example, in the sample collection step, a human swab is used to collect body fluids and then the swab is placed in a test tube containing a specific solution. In this way, various foreign substances are separated by a specific solution, along with the sample on the swab, and in the process, the protein or phospholipid surrounding the cell nucleus is destroyed, and a gene in the nucleus is obtained.

Subsequently, a collection step is performed in which the genes dispersed in a specific solution in the test tube are collected using magnetic beads. Magnetic beads used in the collecting step are known in the art, and are composed of a ball-shaped magnetic material (for example, ferrous metal), a coating layer made of plastic or glass surrounding the magnetic material, and a silica coating layer covering the coating layer. This silica coating layer has the property of attracting and attracting genes. Therefore, when a large number of magnetic beads are added to an in vitro solution, genes dispersed in a specific bath solution adhere to the surface of the magnetic beads (silica coating layer).

In this state, a bar magnet, more specifically an electromagnet, is placed in a test tube and generates a magnetic force, and the magnetic material in the magnetic bead gathers and sticks around the bar magnet in response to the magnetic force of the bar magnet.

Subsequently, a washing step is performed to remove impurities other than genes stuck to the magnetic beads. The washing step is typically carried out twice, but there is no limit to the number of runs.

In addition, the washing step is carried out in a vessel containing a washing buffer (washing liquid or washing liquid).

More specifically, in the first washing step, a bar magnet in which a plurality of magnetic beads are stuck is put in a test tube containing a washing buffer, and a current is cut off to lose the magnetic force of the electromagnet.

In this way, a number of magnetic beads that have stuck to the bar magnet are released from the bar magnet and dispersed in the wash buffer in the test tube. By leaving it in this state for a certain period of time, impurities are settled or separated. Thereafter, a current is applied to the demagnetized bar magnet to remagnetize the bar magnet so that the magnetic beads to which the gene is adsorbed are attached to the bar magnet.

Then, the bar magnet, which has a large number of magnetic beads, is transferred to the second cleaning step, and as described in the first cleaning step, the magnetic force of the bar magnet is removed and the magnetic force of the bar magnet is restored again after a certain time. The second wash is performed.

Next, a gene eluting step of separating the gene from the rod magnet is performed. This gene elution step is performed using an elution buffer (eluent) that prevents the gene from sticking to the magnetic beads.

More specifically, the magnetic beads adhering to the rod magnets are dispersed in the ion buffer by inserting the bar magnets transferred from the previous washing step into the gene eluting step into the test tube containing the illu- tion buffer and blocking the current. After a certain time has elapsed in this state, magnetizing the current by flowing the rod magnet again, the magnetic beads that have fallen off the gene will stick to the rod magnet.

However, the above-described conventional genetic testing method is complicated to separate and elute the gene from the collected sample, and the size of the device for performing this is so large that the genetic test can be performed if the device does not move to the place where it is installed. There is no problem.

Meanwhile, as a result of examining the prior art related to the present invention, a number of patent documents have been searched, and some of them are introduced as follows.

First, the Republic of Korea Patent Publication No. 10-1707987 (name: the continuous quantitative divider and the continuous gene extraction-amplification equipment and the continuous gene extraction-amplification method including the divider) is connected to the external actuator can be connected A volume variable part formed at one end,

A pipette portion connected to the other end of the volume variable portion so as to suck or discharge a predetermined amount of liquid in response to the volume change amount of the volume variable portion generated when the external actuator is driven while being coupled to the connection end of the volume variable portion; And a locking jaw formed on the outer circumferential surface of the pipette part, and when the external actuator descends to the test tube containing the liquid, the locking actuator formed on the outer circumferential surface of the pipette part is caught by the test tube edge and the external actuator is reduced in volume An automatic quantitative divider characterized in that the liquid is aspirated when the gas is raised, a sample kit seating unit in which a sample kit capable of performing gene extraction through an automated series of processes, a PCR amplifier for performing gene amplification, A continuous gene extraction-amplification equipment comprising: a waste disposed between the sample kit seat and the PCR amplifier; and a rod magnet moving rail for transferring the bar magnet from the sample kit seat to the PCR amplifier according to a given processing recipe. As the sample kit, the contamination chamber is stored in the contamination prevention tip for fitting to the end of the bar magnet Tip storage unit; A magnetic bead storage unit having a plurality of magnetic beads having magnetic beads that can stick to the rod magnets and silica coating layers to which the genes can be adsorbed; A sample sample storage unit capable of injecting a liquid sample in which genes and inevitable impurities are mixed; Washing buffer storage for relatively high impurity concentration while increasing the gene concentration; An illution buffer storage unit configured to store an illution buffer for containing only genes; And an automatic metering dispenser storage unit having a built-in metering dispenser disposed at intervals from each other, wherein the disposal unit separates and removes (drops) the contamination prevention tip inserted into the bar magnet and the end of the bar magnet. By using the continuous gene extraction-amplification equipment and the continuous gene extraction-amplification equipment, characterized in that the notch is formed on one side to enable, characterized in that it automates the entire process from gene extraction to gene amplification A continuous gene extraction-amplification method is disclosed.

Next, the Republic of Korea Patent Publication No. 10-1741639 (name: gene or biological material separation and test kit) is formed with an opening 11 is formed on one side of the lower end and the support 10 is formed with a coupling portion on the top; The support 10 is provided with one lysing buffer receiving portion 15c and two washing buffer containing portions 15b filled with the lysing buffer containing the sample, and a hole 15a for installing the elution buffer container. Specimen case 15 coupled to the opening 11 of the; A lower case 20 coupled to an upper side of the support 10 and provided with a locking jaw 21 having a fixing groove 22 formed at an interval of 90 degrees on an upper portion thereof; An elastic member 45 having a protrusion 46 coupled to the locking jaw 21 at a lower end thereof is formed at intervals of 90 degrees, and has a cylindrical rotating member rotatably coupled to an upper side of the lower case 20. 40); An elastic part 61 having a hook 62 coupled to a lower portion of the protrusion 46 of the rotating member 40 in the fixing groove 22 of the lower case 20 is formed at a lower end at an interval of 90 degrees. A primary button 60 disposed inside the rotating member 40; A tip member having a test tube-shaped tip 81 inserted into each buffer of the specimen case 15 and supported by a tip holder 85 coupled to the through portion 63 of the primary button 60 ( 80); A through hole 32 is formed inside the lower case 20, supported by the support 10, and a tip 81 of the tip member 80 is inserted into an upper surface thereof. An inner case 30 having a spring sheet 33 on which the outer spring 35 elastically supporting) is seated; The magnetic member 51 inserted into the tip 81 of the tip member 80 is installed in the lower body 52 and the support rod 53 is formed on the upper center, and the lower body 52 is the inner spring ( A body portion 50 elastically supported by 55; A secondary button (75) installed in the through portion (63) of the primary button (60) to lift the body portion (50); Genetic or biological, characterized in that it comprises; button cover 70 is coupled to the upper side of the primary button 60, the coupling protrusion 71 is coupled to the top of the secondary button 75 formed on the bottom surface Disclosed is a material separation and test kit.

Next, the Republic of Korea Patent Publication No. 2002-0029477 (name: gene extraction method in one step) in a portion of the tissue or cells of the animal in the lysis buffer reagent and crushed to crush the lysis buffer reagent for a predetermined time After the gene was extracted through centrifugation, the PCR was performed immediately using a predetermined amount of the supernatant, thereby reducing the time and simplifying the experimental method, thereby enabling a large-scale experiment. The extraction method is described.

Next, the Republic of Korea Patent Publication No. 10-1025135 (name: automatic purification device, multi-well plate kit and a method for extracting nucleic acids from biological samples) using magnetic particles reversibly bind to the magnetic particles from a plurality of biological samples An apparatus for separating a target material, comprising: a pipette block mounted on at least two rows such that a plurality of pipettes are separable, and for sucking and discharging a biological sample including a target material on each of the plurality of pipettes; A fixed body supporting the pipette block; A magnetic field applying unit for applying and releasing a magnetic field to each row of pipettes mounted on the pipette block; Pipette block vertical movement means for moving the pipette block in the vertical direction; Disclosed is an automatic purifying apparatus comprising a pipette block forward and backward movement means for moving the pipette block forward and backward.

Republic of Korea Patent Publication No. 10-1707987

The present invention has been made in order to solve the above problems of the prior art, is formed of a simple structure is easy to carry and can perform a genetic test for the sample collected quickly and anyone can easily use the extraction device The purpose is to provide.

In addition, it aims to provide an extraction device that can be used quickly and easily and economically by automating the whole process.

The present invention has been made to solve the above-mentioned problems of the prior art, is formed of a simple structure is easy to carry and can carry out a genetic test on the collected sample quickly and can be easily used by anyone and automate the whole process The purpose of the present invention is to provide an extraction apparatus that can be used quickly and easily and economically. As a means for achieving the above object, a buffer containing portion filled with a sample buffer solution and a washing buffer solution are filled. A sample case including a washing buffer receiving portion and an illution buffer receiving portion filled with the Illution buffer liquid; a plurality of test tube-shaped tips inserted into each receiving portion of the sample case are provided with a tip holder supporting the upper portions of the plurality of tips A tip member having a plurality of magnets inserted into the tip member A body portion having a member; And a driving unit for moving the tip member and the body portion coupled to each other in the up, down, left, and right directions to be immersed in each receiving portion of the specimen case.

The apparatus may further include a mixing prevention plate disposed between the body portion and the sample case to prevent the buffer solution buried on the surface of the body portion when the body portion moves to the sample case.

In addition, the mixing plate is characterized in that it is repeatedly moved in the left, right direction along the tip member and the body portion coupled to each other.

The present invention has the following effects.

First, since it is formed in a simple structure, it is easy to carry and of course, the whole process for genetic testing can be performed sequentially from nucleic acid extraction process to mixing process of nucleic acid and sample by simple manipulation. There is an effect that it is possible to check the results quickly by performing the test.

Second, since the magnetic member is inserted into each buffer in a state accommodated in the tip, contamination of the magnetic member is prevented.

Third, the effect buffer container can be implemented in various forms, so that the appropriate type of buffer buffer can be applied as needed.

Fourth, there is an effect that can be used quickly and easily and economically by automating the whole process.

1 to 3 show the overall appearance of the extraction apparatus according to the present invention.
4 is a view showing the appearance of the housing is omitted in the extraction device shown in FIG.
5 is a front view of the extraction apparatus shown in FIG.
6 to 7 are views for explaining a driving unit that is the main part of the present invention.
8 is a view for explaining the concept of a specimen case.
9 is a view showing the positional relationship between the specimen case, the tip member, the body portion, the mixing plate.
10 is a plan view of the extraction apparatus shown in FIG.

Objects, features, and advantages of the present invention described above will become more apparent through the following embodiments in conjunction with the accompanying drawings. The following specific structures or functional descriptions are only illustrated for the purpose of describing the embodiments according to the inventive concept, and the embodiments according to the inventive concept may be implemented in various forms, and the embodiments described herein. It should not be construed as limited to the examples.

Since embodiments according to the concept of the present invention can be variously modified and have various forms, specific embodiments are illustrated in the drawings and will be described in detail herein. However, this is not intended to limit the embodiments in accordance with the concept of the present invention to a specific disclosed form, it should be understood to include all modifications, equivalents, and substitutes included in the spirit and scope of the present invention.

Terms such as first and / or second may be used to describe various components, but the components are not limited to the terms. The terms are only for the purpose of distinguishing one component from other components, for example, without departing from the scope of the rights according to the inventive concept, and the first component may be called a second component, and similar For example, the second component may also be referred to as a first component.

When a component is referred to as being "connected" or "connected" to another component, it may be directly connected or connected to that other component, but it may be understood that other components may be present in the middle. Should be. On the other hand, when a component is said to be "directly connected" or "directly connected" to another component, it should be understood that no other component exists in the middle. Other expressions for describing relationships between components, such as "between" and "immediately between" or "adjacent to" and "directly adjacent to", should be interpreted as well.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. The terms "comprise" or "having" herein are intended to indicate that there is a feature, number, step, action, component, part, or combination thereof that is practiced, and that one or more other features or numbers, It is to be understood that it does not exclude in advance the possibility of the presence or addition of steps, actions, components, parts or combinations thereof.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art. Terms such as those defined in the commonly used dictionaries should be construed as having meanings consistent with the meanings in the context of the related art, and are not construed in ideal or excessively formal meanings unless expressly defined herein. Do not.

1 to 3 is a view showing the overall appearance of the extraction apparatus according to the invention, Figure 4 is a view showing the appearance of the housing is omitted in the extraction device shown in Figure 3 and Figure 5 is shown in Figure 4 6 to 7 are views for explaining a driving unit which is the main part of the present invention, and FIG. 8 is a view for explaining the concept of a sample case, and FIG. 9 is a sample case, a tip member, It is a figure which shows the positional relationship between a body part and a mixing prevention plate.

The present invention is formed of a simple structure is easy to carry and can carry out a genetic test on a sample collected quickly and can be easily used by anyone, and it is fast and easy to use and economical by automating the whole process. have.

Extraction apparatus according to the present invention is largely as shown in the accompanying drawings to implement the above features, the buffer containing portion filled with the lysis buffer liquid containing the sample, the washing buffer receiving portion and the washing buffer liquid is filled with A sample case 20 including an illu- tion buffer receiving portion filled with a buffer liquid, and a plurality of test tube-shaped tips inserted into each receiving portion of the sample case 20 are provided, and a tip holder supporting the upper portions of the plurality of tips is provided. The specimen 30 has a tip member 30, a body portion 40 having a plurality of magnetic members inserted into the tip member 30, and the tip member 30 and the body portion 40 coupled to each other. It characterized in that it comprises a drive unit 50 for moving in the up, down, left, right direction to be immersed in each receiving portion of the case 20.

On the other hand, the extraction apparatus according to the present invention may be provided with a housing 10 to configure the appearance of the extraction apparatus, the sample case 20, the tip member 30, the body described above in the inner space of the housing 10 Members such as the part 40 and the driving part 50 may be disposed in place.

More specifically, the tip member 30 is configured such that when the magnetic member is inserted into the tip, a plurality of magnetic beads included in the lysis buffer and having a silica coating layer formed on the outer surface thereof are adsorbed on the outer surface of the tip. This is for the magnetic member to absorb and transfer the magnetic beads to which the genetic component of the sample is attached by using the magnetic force of the magnetic member without directly contacting the lysis buffer including the sample.

Next, the driving unit 50 is responsible for providing a driving force so that the tip member 30 and the body 40 in the upper portion of the sample case 20 can move freely in the up, down, left, and right directions. The drive unit 50 is preferably implemented so as to raise and lower the tip member 30 and the body portion 40, respectively.

To this end, the driving unit 50 includes a first ball screw 51 extending in the up and down directions, a first rotation motor 52 for rotating the first ball screw 51, and the first ball screw 51. It may include a first guide block 53 which moves in the up, down direction along the longitudinal direction of the.

Here, the tip member 30 is installed in the first guide block 53, and the tip member 30 is interlocked in the up and down directions along the first guide block 53 moving upward and downward. .

In addition, the second ball screw 54 installed in parallel with the first ball screw 51, the second rotation motor 55 for rotating the second ball screw 54, the second ball screw 54 It may further include a second guide block 56 to move in the up, down direction along the longitudinal direction of the).

Here, the body portion 40 is installed in the second guide block 56, the body portion 40 is interlocked in the up and down direction along the second guide block 56 moving in the up and down direction do.

That is, the driving unit 50 moves the tip member 30 and the body portion 40 separately within the housing 10 to perform the mutual coupling action of the body portion 40 and the tip member 30. .

On the other hand, the driving unit 50 may move the tip member 30 and the body portion 40 in the left and right directions at the same time. To this end, the drive unit 50 may further include a translation means.

In this case, the tip member 30 and the body portion 40 are fixed on the mounting bracket 60, the translation means translate the mounting bracket 60 as a whole, the tip member 30 and the body portion 40 Illustrates an example in which the left and right translations are performed.

The translation means according to the present embodiment includes a main motor, an interlocking pulley connected to a belt at an output end of the main motor, and a mounting bracket 60 is fixed on the belt.

That is, when the main motor is driven, the mounting bracket 60 translates in the left and right directions between the output terminal of the main motor and the interlocking pulley.

Accordingly, the tip member 30 and the body portion 40 respectively mounted on the mounting bracket 60 may be repeatedly moved in the left and right directions.

Hereinafter, the operation of the extraction apparatus according to the present invention. In this action, a method of extracting a gene of a sample using the extracting apparatus according to the present invention will be described as an example.

First, the process starts with taking a sample from a test object.

For example, in the sample collection step, a human swab is used to collect body fluids and then the swab is placed in a test tube containing a specific solution.

In this way, various foreign substances are separated by a specific solution together with the sample on the swab, and in this process, the protein or phospholipid surrounding the cell nucleus is destroyed, and a gene in the nucleus is obtained.

Subsequently, a collection step is performed in which the genes dispersed in a specific solution in the test tube are collected using magnetic beads. Magnetic beads used in the collecting step are known in the art, and are composed of a ball-shaped magnetic material (for example, ferrous metal), a coating layer made of plastic or glass surrounding the magnetic material, and a silica coating layer covering the coating layer. This silica coating layer has the property of attracting and attracting genes. Therefore, when a large number of magnetic beads are added to an in vitro solution, genes dispersed in a specific bath solution adhere to the surface of the magnetic beads (silica coating layer).

Next, the magnetic beads in which the genes are attached are mixed with the lysis buffer solution and stored in the lys buffer buffer of the sample case 20 according to the present invention.

In this case, the sample case 20 is preferably implemented to be detachable from the housing 10, and at the same time, the buffer buffer used in each step is filled in the washing buffer accommodating portion and the elution buffer accommodating portion.

In this state, the tip member 30 and the body portion 40 are lowered toward the lysis buffer accommodating portion so as to completely immerse the tip member 30 in the sis buffer accommodating portion. Then, the magnetic bead in the magnetic bead gathers and sticks around the tip member 30 in response to the magnetic force of the magnetic member of the body portion 40.

Subsequently, a washing step is performed to remove impurities other than genes stuck to the magnetic beads. The washing step is usually performed twice, but there is no limit to the number of times.

The washing step may be performed by immersing the magnetic beads in the washing buffer receiving portion of the sample case 20.

To this end, the tip member 30 and the body portion 40 immersed in the lysis buffer receiving portion is raised to move to the right. At this time, the extraction device according to the present invention, the tip member (30) to prevent the buffer liquid on the surface of the tip member 30 to fall to the sample case 20 when the body portion 40 and the tip member 30 is moved. 30 may further include a mixing prevention plate 70 disposed between the sample case 20 and the mixing prevention plate 70 along the tip member 30 and the body 40 coupled to each other. It is implemented to move repeatedly in the left and right directions.

That is, when the tip member 30 and the body portion 40 are lifted from the lysis buffer receiving portion, the anti-mixing plate 70 immediately moves downward of the tip member 30 to flow the buffer liquid flowing from the tip member 30. I receive it.

In this state, the tip member 30 and the body portion 40 move upward of the washing buffer accommodating portion and are immersed toward the washing buffer accommodating portion when the anti-mixing plate 70 is removed.

The tip member 30 to which the plurality of magnetic beads are stuck is placed in the receiving portion containing the washing buffer and the current is interrupted to lose the magnetic force of the body portion 40.

In this way, a plurality of magnetic beads stuck to the tip member 30 are released from the tip and distributed to the washing buffer in the washing buffer receiving portion. By leaving it in this state for a certain period of time, impurities are settled or separated. Subsequently, a current flows through the body portion 40 from which the magnetic force is removed, thereby magnetizing the body portion 40 again, thereby causing the magnetic beads to which the gene is adsorbed to adhere to the tip member 30.

Next, a gene eluting step of separating the genes is performed. This gene elution step is performed using an elution buffer (eluent) that prevents the gene from sticking to the magnetic beads.

The illution buffer is filled in the illution buffer accommodating part of the sample case 20. For this purpose, the tip member 30 and the body part 40 which are immersed in the washing buffer accommodating part are raised to position the upper part of the elution buffer accommodating part. Immerse in the solution buffer receptacle.

Next, by blocking the current of the body portion 40, the magnetic beads adhering to the tip member 30 are dispersed in the illusion buffer. After a certain time has elapsed in this state, when the current flows through the body 40 again and magnetizes, the magnetic beads from which the genes have fallen out adhere to the surface of the tip member 30.

As a result, only the pure gene to be extracted is retained in the elution buffer accommodating part, and the sample case 20 is removed from the housing 10 to be used for experiments using other genes.

In summary, the extraction apparatus according to the present invention has a simple structure and is easy to carry, and can be used to perform the overall process for genetic testing sequentially from a nucleic acid extraction process to a nucleic acid and sample mixing process by a simple operation. This is an easy and simple on-site genetic test that has the effect of being able to quickly verify the results.

In addition, since the magnetic member is inserted into each buffer in a state accommodated in the tip, the magnetic member is prevented from being contaminated.

In addition, according to the separation and test kit of the gene or biological material of the present invention, by implementing the elution buffer container in various forms, there is an effect that can be applied to the appropriate type of elution buffer container as needed.

Embodiments of the present invention described above are merely exemplary, and those skilled in the art will appreciate that various modifications and equivalent other embodiments are possible therefrom. Therefore, it will be understood that the present invention is not limited to the forms mentioned in the above detailed description. Therefore, the true technical protection scope of the present invention will be defined by the technical spirit of the appended claims. It is also to be understood that the present invention includes all modifications, equivalents and substitutes within the spirit and scope of the invention as defined by the appended claims.

10: housing
20: sample case
30: tip member
40: body part
50: drive unit
60: mounting bracket
70: mixing plate

Claims (3)

A sample case including a buffer accommodating portion filled with a lysine buffer liquid containing a sample, a washing buffer accommodating portion filled with a washing buffer liquid, and an elution buffer accommodating portion filled with the washing buffer liquid;
A tip member having a plurality of test tube-shaped tips inserted into each receiving portion of the specimen case and having a tip holder supporting the plurality of tip upper portions;
A body part having a plurality of magnetic members inserted into the tip member; And
And a driving unit for moving the tip member and the body unit which are coupled to each other in up, down, left, and right directions to be immersed in the respective receiving portions of the specimen case. The method according to claim 1,
And a mixing prevention plate disposed between the body portion and the sample case to prevent the buffer solution from the surface of the body portion falling to the sample case when the body portion is moved. The method according to claim 2,
The anti-mixing plate is a biological material extraction device, characterized in that for repeatedly moving in the left, right direction along the tip member and the body portion coupled to each other.

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